Study On The Mechanism Of The Traditional Chinese Medicine Qiliqiangxin In Reducing The Right Heart Failure Caused By Pulmonary Hypertension In Rats

Objective:The aim was to observe the therapeutic effect of traditional Chinese medication qiliqingxin(QLQX)on the rats with right ventricular failure due to pulmonary arterial hypertension(PAH-RVF)induced by monocrotaline(MCT).Methods:Male SD rats were randomly divided into negative control group(Vehicle+Saline),PAH-RVF group(MCT+Saline)and QLQX treatment group(MCT+QLQX).PAH-RVF model was induced by of a single intraperitoneal injection with 60mg/kg MCT.Rats were weighted weekly.Two weeks later,rats from MCT+QLQX group recived lg/kg QLQX by intragastric administration daily.Four weeks later,echocardiography and right heart catheterization were carried out and right ventricular systolic pressure(RVSP)was recorded.The right heart index,right heart weight/body weight(RW/BW),heart weight/body weight(HW/BW),lung weight/body weight(LW/BW)were calculated.H&E and masson staining were performed with the heart and lung tissues and histological observation was carried out.RT-PCR was used to detect cardiac atrial natriuretic peptide(ANP),brain natriuretic peptide(BNP),CollagenI,CollagenⅢ,peroxisome proliferator activated receptor γ(PPARγ).The apoptosis related proteins such as B-cell Lymphoma 2(Bcl2),Bcl2 associated X(Bax),cleaved cysteinyl aspartate specific proteinase(Cleaved-caspase3)/Caspase3,p27 and fibrosis related proteins like tissue inhibitor of metalloproteinase 3(TIMP3),transforming growth factor-beta(TGF-β),matrix metallopeptidase 2(MMP2),matrix metallopeptidase 9(MMP9),as well as common proteins involved in signaling pathways such as peroxisome proliferator activated receptor gamma(PPARγ),peroxisome proliferator activated receptor gamma coactivator-1alpha(PGC-la),phosphorylated protein kinase B 473(p-Akt473)/total protein kinase B(t-Akt),phosphorylated extracellular signal regulated kinases(p-Erk)/total extracellular signal regulated kinases(t-Erk),phosphorylated endothelial nitric oxide synthase1177(p-eNOS1177)/total endothelial nitric oxide synthase(t-eNOS)in heart and(or)lung were detected by western blot.Results:After received an intraperitoneal MCT injection,the rats’ weight lost while the RVSP,RW/BW,HW/BW,LW/BW gained.Hearts were infiltrated with inflammatory cells and cardiomyocytes disarranged.The thickness of small pulmonary artery medial wall and its percentage of vessel diameter as well as the fibrosis increased.The protein of p27,PPARγ,PGC-1α,Bax/Bcl2,p-eNOS1177/t-eNOS decreased while TGF-P,MMP2,MMP9,TIMP3,p-Erk/t-Erk increased in lung.Besides,the cardiac fibrosis was aggravated.The cardiac mRNA of ANP,BNP,CollagenI,CollagenⅢ increased and PPARγ reduced.The protein of PPARγ,PGC-1α was down-regulated;TGF-β,MMP2,MMP9,Bax/Bcl2,p-Erk/t-Erk,Cleaved-caspase3/Caspase3 was up-regulated;p-Akt473/t-Akt didn’t change significantly in heart.With QLQX treatment and compared with MCT+Saline group,the body weight of the rats didn’t change significantly while the RVSP,RW/BW,HW/BW,LW/BW decreased.There were less inflammatory cells and the cardiomyocytes architecture was preserved.The thickness of small pulmonary artery medial wall and its percentage of vessel diameter as well as its fibrosis were closer to the normal.The protein expression in lung such as p27,PPAR γ increased eased and TGF-β,MMP2,MMP9 decreased while p-eNOS1177/t-eNOS,p-Erk/t-Erk had no significant changes.The cardiac fibrosis and the mRNA levels of ANP,BNP,CollagenⅠ,CollagenⅢ reduced.However,PPARγ mRNA increased.At protein level,the cardiac PPAR y increased;TGF-β,MMP2,MMP9,Bax/Bcl2,Cleaved-caspase3/Caspase3 decreased and PGC-1α,p-Erk/t-Erk,p-Akt473/t-Akt didn’t change significantly.Conclusions:1.Qiliqiangxin can reduce right ventricular systolic pressure and alleviate pulmonary remodeling in PAH-RVF models.2.Qiliqiangxin is capable of reducing the apoptosis and fibrosis of right ventricular and improving cardiac remodeling in PAH-RVF models.Objective:To explore the molecular mechanisms of qiliqiangxin on rats with right ventricular failure due to pulmonary arterial hypertension.Methods:1.Male SD rats were randomly divided into Vehicle+Saline group,Vehicle+QLQX group,MCT+Saline group,MCT+QLQX group,MCT+QLQX+PPARγ agonist group and MCT+QLQX+PPARγ inhibitor group.PAH-RVF model were induced by of a single intraperitoneal injection with 60mg/kg MCT.After 2 weeks,rats received QLQX and PPARγ agonist/inhibitor.At the end of four weeks,RVSP,RV index,RW/BW,HW/BW,LW/BW were determined.H&E and masson staining were performed with the heart and lung tissues and histological observation was carried out.Western blot was used to detect PPARγ,Bax,Bcl2,TGF-β in heart and lung tissues.2.Primary cardiomyocytes were separated from neonatal rats and were stimulated by PE for myocardial hypertrophy model.Given QLQX and PPARγ agonist/inhibitor,RT-PCR was used to detect ANP and BNP.The size of cardiomyocytes was measured by immunefluorescence.Pulmonary artery smooth muscle cells were separated from SD rats and were stimulated by PDGF-BB.Given QLQX and PPARγ agonist/inhibitor,the EdU assay was used for cell proliferation.Results:1.PPARγ agonist didn’t provide additional improvements in the presence of QLQX while PPARγ inhibitior reduced the beneficial effects of QLQX on PAH-RVF including RVSP,RV index,RW/BW,HW/BW,LW/BW,pulmonary arterioles medial wall thickness and its percentage accounted for diameter,heart and lung fibrosis,the protein of TGF-β and Bax/Bcl2.2.PE increased cardiomyocytes’ sizes,the mRNA levels of ANP and BNP;QLQX reduced cardiomyocytes’ sizes,ANP and BNP;PPARγ agonist didn’t provide additional improvements in the presence of QLQX and PPARγ inhibitior reduced the beneficial effects of QLQX on cardiomyocytes.Pulmonary artery smooth muscle cells were stimulated by PDGF-BB.EdU positive cells were decreased by QLQX.PPARγ agonist didn’t provide additional improvements in the presence of QLQX.PPARγ inhibitior reduced the effects of QLQX on PASMCs.Conclusions:1.QLQX attenuates the rats with right ventricular failure duo to pulmonary arterial hypertension by increasing PPARγ levels.2.QLQX reduces myocardial hypertrophy induced by PE and inhibits proliferation of pulmonary artery smooth muscle cells induced by PDGF-BB through PPARγ pathway.